Discovery, recognized antigenic structures, and evolution of cross-serotype broadly neutralizing antibodies from porcine B-cell repertoires against foot-and-mouth disease virus.

It is a great challenge to isolate the broadly neutralizing antibodies (bnAbs) against foot-and-mouth disease virus (FMDV) due to its existence as seven distinct serotypes without cross-protection. Here, by vaccination of pig with FMDV serotype O and A whole virus antigens, we obtained 10 bnAbs against serotypes O, A and/or Asia1 by dissecting 216 common clonotypes of two serotype O and A specific porcine B-cell receptor (BCR) gene repertoires containing total 12720 B cell clones, indicating the induction of cross-serotype bnAbs after sequential vaccination with serotypes O and A antigens. The majority of porcine bnAbs (9/10) were derived from terminally differentiated B cells of different clonal lineages, which convergently targeted the conserved "RGDL" motif on structural protein VP1 of FMDV by mimicking receptor recognition to inhibit viral attachment to cells. Cryo-EM complex structures revealed that the other bnAb pOA-2 specifically targets a novel inter-pentamer antigen structure surrounding the viral three-fold axis, with a highly conserved determinant at residue 68 on VP2. This unique binding pattern enabled cross-serotype neutralization by destabilizing the viral particle. The evolutionary analysis of pOA-2 demonstrated its origin from an intermediate B-cell, emphasizing the crucial role of somatic hypermutations (SHMs) in balancing the breadth and potency of neutralization. However, excessive SHMs may deviate from the trajectory of broad neutralization. This study provides a strategy to uncover bnAbs against highly mutable pathogens and the cross-serotype antigenic structures to explore broadly protective FMDV vaccine. Author summary: The scarcity of broadly neutralizing antibodies (bnAbs) in the body significantly hinders their discovery and elucidation of the conserved antigenic structure of highly mutable pathogens. Foot-and-mouth disease virus (FMDV) severely infects pigs and exists as seven distinct serotypes without cross-protection. In this study, we present a strategy for rapid discovery of cross-serotype bnAbs by constructing of specific B-cell repertoires and exploring of common clonotypes. We originally isolated porcine bnAbs against FMDV serotypes O, A and/or Asia1, and proved for the first time that receptor binding region in VP1 can induce the production of cross-serotype bnAbs. The evolution of cross-serotype bnAbs reveals how somatic hypermutations balance the breadth and potency of neutralization. FMDV bnAbs employ dual neutralization mechanisms to neutralize viruses via receptor mimicry and destabilizing the viral particles. This study provides valuable guidelines that facilitate the isolation of bnAbs against the highly variable viruses and aid in designing a universal FMDV vaccine. [ABSTRACT FROM AUTHOR]